Acidic polysaccharide CP-2 from Dioscoreae Rhizoma ameliorated acute alcoholic liver injury through the gut-liver axis and AMPK/PPAR pathway

3.0 科研～小助 2025-08-30 7 4 18.74MB 14 页 1知币

侵权投诉

Acidic polysaccharide CP-2 from Dioscoreae Rhizoma ameliorated acute

alcoholic liver injury through the gut-liver axis and AMPK/PPAR pathway

Yujun Xie

, Mingxing Tao

, Xiaodong Yan

, Xinxin Fan

a,b

, Alamusi Bayoude

, Yu Lu

Shuangli Zhao

, Boyang Yu

a,*

, Renshi Li

a,*

Jiangsu Provincial Key Laboratory of TCM Evaluation and Translational Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing

211198, China

Division of Pharmaceutical Cell Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 8128582, Japan

ARTICLE INFO

Keywords:

Dioscoreae Rhizoma polysaccharide

Acute alcoholic liver injury

Gut-liver axis

ABSTRACT

Dioscoreae Rhizoma polysaccharides exhibit gastrointestinal protective properties, yet their efcacy against

acute alcoholic liver injury (AALI) remains unexplored. This study identies a novel acidic heteropolysaccharide

(CP-2, Mw =8.4 ×10

kDa) with galactose/galacturonic acid dominance and delineates its multimodal hep-

atoprotective mechanisms. In AALI mice, CP-2 attenuated liver injury by enhancing ADH and ALDH activities

while restoring redox balance via SOD/CAT activation and MDA reduction, and suppressed inammation by

inhibiting IL-1β, IL-6, and TNF-

levels. Gut-liver axis modulation was achieved through intestinal barrier

reinforcement (ZO-1, Occludin, Claudin-1) and microbiota rebalancing. CP-2 could reduce gram-negative bac-

teria ([Ruminococcus]_ torques_ group and Escherichia- Shigella) and Proteobacteria abundance while enriching

Bacteroides and Akkermansia abundance, which collectively suppressed serum LPS level. In addition, CP-2 could

activate the AMPK/PPAR signaling pathway to reduce the production of fatty acids and promote their degra-

dation in AALI. CP-2 can improve AALI by adjusting the composition of gut microbiota, repairing intestinal

barrier function, decreasing systemic inammation and oxidative reactions, and regulating the AMPK/PPAR

pathway. Our ndings unveil CP-2 as a prebiotic candidate for AALI intervention and may advance functional

food development for alcohol-related hepatopathies.

1. Introduction

Alcohol consumption constitutes a widespread global public health

concern. According to recent WHO data, approximately 2.3 billion in-

dividuals consume alcohol, with an estimated 283 million (237 million

males and 46 million females) engaging in hazardous drinking patterns

[1]. Alcohol-induced liver injury, especially acute alcoholic liver injury

(AALI), ranks among the most prevalent alcohol-associated pathologies

worldwide [2]. Without timely and effective intervention, AALI pro-

gresses to advanced stages of alcoholic liver disease, culminating in

irreversible complications such as cirrhosis and brosis [3]. Conse-

quently, developing strategies to mitigate AALI has emerged as a critical

priority in public health research.

The pathogenesis of AALI may be elucidated through the gut-liver

axis, a bidirectional relationship between hepatic function and intesti-

nal homeostasis. Chronic alcohol intake disrupts gut microbiota equi-

librium, compromises intestinal barrier integrity, and elevates intestinal

permeability [4]. These alterations facilitate the translocation of lipo-

polysaccharides (LPS) from gram-negative bacteria into portal circula-

tion, triggering Kupffer cell activation and subsequent pro-inammatory

cytokine cascades that exacerbate hepatic injury [5]. Alcohol is mainly

absorbed by the intestinal canal for metabolism in liver, increasing ROS

release and a concomitant weakening of the antioxidant defense system.

This results in unbalanced oxidative status, which in turn gives rise to

oxidative stress in the liver [6]. Alcohol ingestion can also lead to dis-

turbances in liver lipid metabolism through fats synthesis increasing and

the fatty acids oxidation reducing, ultimately results in an excessive

lipids storage, a condition termed hepatic steatosis [7]. It is generally

accepted that an imbalanced gut microbiota, enhanced gut perme-

ability, hepatic inammation and oxidative stress, as well as lipid

deposition, are involved in AALI development [8]. Current therapeutic

interventions for alcohol-related hepatotoxicity, including anti-

inammatory agents (e.g., diammonium glycyrrhizinate), antioxidants

(e.g., glutathione), and hepatoprotective compounds (e.g.,

* Corresponding authors.

E-mail addresses: boyangyu59@163.com (B. Yu), li-renshi@cpu.edu.cn (R. Li).

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

https://doi.org/10.1016/j.ijbiomac.2025.143145

Received 20 January 2025; Received in revised form 31 March 2025; Accepted 12 April 2025

International Journal of Biological Macromolecules 310 (2025) 143145

Available online 13 April 2025

ursodeoxycholic acid), demonstrate limited efcacy and carry risks of

adverse effects, underscoring the need for safer alternatives. This ther-

apeutic gap has intensied interest in natural bioactive compounds with

multimodal hepatoprotective properties.

Polysaccharides represent a promising class of phytochemicals

exhibiting potent antioxidant, anti-inammatory, and immunomodula-

tory activities [9]. Some polysaccharides extracted from Angelica sinensis

[10], Enteromorpha prolifera [11], and Dendrobium ofcinale [12]

demonstrated anti-AALI activity. Dendrobium huoshanense poly-

saccharides could mitigate early-stage steatosis and inammation in

AALI by modulating the fatty acid metabolism pathway [13]. Dioscoreae

Rhizoma (Dioscorea opposite Thunb.), also known as Chinese yam (CY),

has been a resourceful food for gastrointestinal protection on a global

scale [14]. It has boasted numerous biological activities, such as the

strengthening of immunity, gastrointestinal protection, antioxidation,

anti-tumor, and so on [15]. Earlier literature has shown that yam gruel

can regulate hepatic lipid metabolism disorders by activating the

AMPK/ACC/CPT-1 pathway in diabetic rats [16]. Polysaccharides,

which are among the most prevalent active ingredients in CY, have been

shown to reduce oxidative, inammatory, and anti-hyperglycaemic ef-

fects [17]. However, as a widely used resource plant, the hep-

atoprotective effect of Dioscoreae Rhizoma polysaccharide (CP) has not

been specically studied.

This study investigates the therapeutic efcacy of a novel acidic

polysaccharide from Dioscoreae Rhizoma (CP-2) in AALI models.

Through a comprehensive analysis of ethanol metabolism biomarkers,

oxidative-inammatory markers, lipid-regulatory proteins, and gut

microbiota proles, we elucidate CP-2's mechanistic role in hep-

atoprotection. This research could form the basis for the formulation and

use of CP-2 in food and medical applications.

2. Materials and methods

2.1. Materials and reagents

Anhydrous ethanol (analytical grade) was procured from Shanghai

Titan Scientic Co., Ltd. (Shanghai, China). Bifendate pills (batch

number: 230703) were sourced from Beijing Union Pharmaceutical

Factory Ltd. (Beijing, China). DEAE-52, catalase (CAT), malondialde-

hyde (MDA), superoxide dismutase (SOD), and Ethanol Assay Kit were

obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing,

China). Alcohol dehydrogenase (ADH), acetaldehyde dehydrogenase

(ALDH), triglyceride (TG), alanine aminotransferase (ALT), and aspar-

tate aminotransferase (AST) kits were acquired from Nanjing Jiancheng

Bioengineering Institute (Nanjing, China). Lipopolysaccharide (LPS),

cytochrome P450 family 2 subfamily E member 1 (CYP

), Interleukin

1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor-

(TNF-

)

ELISA kits were purchased from Hunan Aifang Biotechnology Co., Ltd.

(Changsha, China).

2.2. The preparation of crude polysaccharide (CCP)

The Chinese yam was supplied by Jiaozuo Mingren Natural Medicine

Co., Ltd. (Jiaozuo, China). CP was prepared via hot water extraction and

ethanol precipitation [18]. Briey, 1,000 g of crushed and sieved raw

materials were mixed with 10 L of distilled water and heated at 100 ◦C

for 1 h. The residue underwent two additional extractions under iden-

tical conditions. The ltrates were then concentrated to 1 L and

precipitated using 80 % ethanol. The precipitate was re-dissolved,

deproteinized using a mixture of chloroform (purity 99.0 %) and n-

butanol (purity 99.5 %) in a 5:1 volume ratio, decolorized by the acti-

vated carbon (Macklin Co., Ltd., Beijing, China), dialyzed (3.5 kDa

MWCO membrane, Beijing Solarbio Science & Technology Co., Ltd.,

Beijing, China), and lyophilized to obtain the crude polysaccharide

(CCP, yield 0.5 %).

2.3. The isolation and purication of CP

CP was subjected to isolation and purication using DEAE-Cellulose

(Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) [19].

Firstly, 50 mg of CCP was dissolved in 1.0 mL of distilled water and

fractionated on the column (50 cm ×2.6 cm). The column was then

eluted with 0, 0.1, 0.3, and 0.5 M NaCl solutions, resulting in the gen-

eration of an elution curve. This curve exhibited three principal peaks,

designated CP-1, CP-2, and CP-3.

2.4. Characterization of CP-2

2.4.1. Chemical composition analysis

Chemical methods were used to determine the total carbohydrate,

protein, and uronic acid content of CP-2 [20–22]. The phenol‑sulfuric

acid method was used to determine the total carbohydrate content of CP-

2. The Coomassie brilliant blue method was used to determine the total

protein content. The m-hydroxydiphenyl colorimetry method was used

to determine the uronic acid content.

2.4.2. CP-2 molecular weight determination

An Agilent 1260 series HPLC equipped with an evaporative light-

scattering detector (ELSD) was used to determine the molecular

weight of CP-2 [21]. Dextran standards of different molecular weights

(China National Institute for the Control of Pharmaceutical and Bio-

logical Products, purity ≥98.0 %) for standard curves. TSK gel GMPWxl

and TSK gel G3000PWxl (Tosoh Corp., Tokyo, Japan) columns were

used for separation at a column temperature of 25 ◦C and a ow rate of

1.0 mL/min. The mobile phase was distilled water using isocratic

elution.

2.4.3. FT-IR spectra determination

CP-2 mixed with potassium bromide powder was used to press mil-

limetre thick sheets. Fourier transform infrared (FT-IR, Shimadzu,

Japan) spectral analysis was performed in the 4000–400 cm

−1

region

[23].

2.4.4. Determination of the composition of CP-2 monosaccharides

CP-2 (5 mg) was dissolved in 2 mL of 3 M triuoroacetic acid, hy-

drolyzed at 120 ◦C for 3 h, and dried with nitrogen gas. The resulting

residue was dissolved in pure water and centrifuged (12,000 rpm, 5

min). Finally, the supernate was used for high-performance anion ex-

change chromatography - pulsed amperometric detector (HPAEC-PAD,

Thermo Fisher ICS5000, USA) analysis [24]. DionexCarbopac™PA20

(150 mm ×3.0 mm) column was used for separation at a column tem-

perature of 30 ◦C and a ow rate of 0.3 mL/min. There were three

mobile phases, including H

O (A), 250 mM NaOH (B), and 500 mM

NaOH and 50 mM NaAc (C). Elution gradient: 98 % A: 2 % B at 0–23

min, 80 % A: 20 % B at 23–33 min, 80 % A: 20 % C at 33–46 min, 20 % A:

80 % C at 46–66 min, 98 % A: 2 % B at 66–80 min. Monosaccharide

standards (BoRui Saccharide Biotech Co. Ltd., purity ≥98.0 %) are used

to determine monosaccharide retention times. Take each mono-

saccharide standard solution and accurately congure the mixed stan-

dard solution. According to the absolute quantitative method, the

content of different monosaccharides was determined and the molar

ratio was calculated according to the molar mass of monosaccharides.

2.5. Animals and treatments

2.5.1. Experimental design

Thirty-six male ICR mice (6 weeks old) were obtained from Yangz-

hou University (Jiangsu, China) and fed according to the procedure of

the Animal Ethics Committee of China Pharmaceutical University

(protocol no. 2023–04-011). Mice were housed at room temperature of

25 ±1 ◦C, relative humidity of 45.0 ±5.0 %, and a 12 h light/dark

cycle.

Y. Xie et al.

International Journal of Biological Macromolecules 310 (2025) 143145

文档加载中……请稍候！
如果长时间未打开，您也可以点击刷新试试。

下载文档到电脑，查找使用更方便

1 知币 4人已下载

立即下载

Acidic polysaccharide CP-2 from Dioscoreae Rhizoma ameliorated acute alcoholic liver injury through the gut-liver axis and AMPK/PPAR pathway.pdf

共14页,预览5页

还剩页未读，继续阅读

Acidic polysaccharide CP-2 from Dioscoreae Rhizoma ameliorated acute alcoholic liver injury through the gut-liver axis and AMPK/PPAR pathway

相关推荐

开通VIP享超值会员特权

作者详情

相关内容

推荐作者

热门标签

举报选择: